钌(Ⅱ)配合物[Ru(dmp)_2(NMIP)](ClO_4)_2抗骨肉瘤HOS细胞的凋亡
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摘要
目的探讨钌(Ⅱ)配合物[Ru(dmp)_2(NMIP)](ClO_4)_2对人骨肉瘤HOS细胞的凋亡作用初步作用机制。方法采用荧光定量PCR检测钌(II)配合物作用于人骨肉瘤HOS细胞后RNA变化,蛋白免疫电泳法检测钌(II)配合物作用于人骨肉瘤HOS细胞后细胞凋亡蛋白变化。结果钌配合物作用于HOS细胞24 h后,提取总基因逆转录后做荧光定量PCR。相比对照组,25、50、100μmol/L钌配合物组Bcl_2的mRNA表达明显降低(分别为0.86±0.02、0.52±0.01、0.4±0.01)(P<0.05),而Bax的mRNA表达明显升高(分别为1.18±0.18、1.7±0.10、1.69±0.10),Caspase3的mRNA表达明显升高(分别为1.22±0.11、1.61±0.12、1.62±0.06)。0、25、50、100μmol/L钌配合物能降低细胞体内Bcl2蛋白表达(分别为0.52±0.03、0.55±0.08、0.39±0.02、0.14±0.01),增加Bax蛋白表达(分别为0.52±0.01、0.60±0.02、0.64±0.03、0.62±0.02),增加Caspase3蛋白表达(分别为0.57±0.01、0.61±0.01、0.68±0.01、0.64±0.01)(P<0.05)。结论钌配合物通过线粒体引起的内源性凋亡通路引起骨肉瘤细胞凋亡。
Objective To investigate the preliminary apoptosis mechanism of Ruthenium(II) complex[Ru(dmp)_2(NMIP)](ClO_4)_2 on human osteosarcoma Hos cells. Methods RNA changes in human osteosarcoma HOS cells,treated with ruthenium(II)complex,were detected by QPCR,and apoptotic protein changes in human osteosarcoma Hos cells,treated with Ruthenium(II) complex,were detected by protein immunoelectrophoresis. Results After 24 hours of ruthenium complexes actingi in HOS cells,the total gene was retrieved and quantified by fluorescence quantitative PCR. Compared with the control group,the expression of Bcl2 in ruthenium complex group decreased significantly(P<0.05),while the expression of Bax and Caspase 3 increased significantly(P<0.05). Ruthenium complexes can decrease the expression of Bcl2 protein and increase the expression of Bax and Caspase 3 protein in cells. Conclusion Ruthenium complex could induce osteosarcoma cell apoptosis through endogenous apoptotic pathway induced by mitochondria.
Objective To investigate the preliminary apoptosis mechanism of Ruthenium(II) complex[Ru(dmp)_2(NMIP)](ClO_4)_2 on human osteosarcoma Hos cells. Methods RNA changes in human osteosarcoma HOS cells,treated with ruthenium(II)complex,were detected by QPCR,and apoptotic protein changes in human osteosarcoma Hos cells,treated with Ruthenium(II) complex,were detected by protein immunoelectrophoresis. Results After 24 hours of ruthenium complexes actingi in HOS cells,the total gene was retrieved and quantified by fluorescence quantitative PCR. Compared with the control group,the expression of Bcl2 in ruthenium complex group decreased significantly(P<0.05),while the expression of Bax and Caspase 3 increased significantly(P<0.05). Ruthenium complexes can decrease the expression of Bcl2 protein and increase the expression of Bax and Caspase 3 protein in cells. Conclusion Ruthenium complex could induce osteosarcoma cell apoptosis through endogenous apoptotic pathway induced by mitochondria.
引文
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[16]华夏,李广兴,刘仲涛,等.人参皂苷Rh2下调Wnt/β-catenin信号通路调控SHG44细胞的增殖和凋亡[J].解剖学研究,2018,40(3):189-192.
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[3] Turer AT,Scherer PE. Adiponectin:mechanistic insights and clinical implications[J].Diabetologia,2012,55:2319-2326.
[4] Lincoln R,Kohler L,Monro S,et al. Exploitation of long-lived 3IL excited states for meta-organic photodynamic therapy:verification in a metastatic melanoma model[J]. J Am Chem Soc,2013,135(45):17161-17175.
[5] Reedijk J. Why does cisplatin reach guanine-N7 with competing S-donor ligands available in the cells[J].Chem Rev,1999,99:2499.
[6] Pettinari R,Pettinari C,Marchetti F,et al. Areneruthenium(II)acylpyrazolonato complexes:apoptosis-promoting effects on human cancer cells[J]. J Med Chem,2014,57(11):4532-4542.
[7] Xu L,Zhong NJ,Xie YY,et al. Synthesis,characterization,in vitro cytotoxicity,and apoptosis-inducing properties of ruthenium(II)complexes[J]. PLoS One,2014,9(5):e96082.
[8] Zhu JW,Liu SH,Zhang GQ,et al. Anticancer activity studies of ruthenium(Ⅱ)complex toward human osteosarcoma cells[J]. J Membrane Biol,2016,249:483-492.
[9] Busto N,Valladolid J,Martínez-Alonso M,et al.Anticancer activity and DNA binding of a bifunctional Ru(II)arene aqua-complex with the 2,4-diamino-6-(2-pyridyl)-1,3,5-triazine ligand[J]. Inorganic Chemistry,2013,52(17):9962-9974.
[10]Gupta RK,Pandey R,Sharma G,et al. DNA binding and anti-cancer activity of redox-active heteroleptic piano-stool Ru(II),Rh(III),and Ir(III)complexes containing 4-(2-methoxypyridyl)phenyldipyrromethene[J]. Inorganic Chemistry,2013,52(7):3687-3698.
[11]Kamatchi TS,Kalaivani P,Poornima P,et al. New organometallic ruthenium(II)complexes containing chelidonic acid(4-oxo-4 H-pyran-2,6-dicarboxylic acid):synthesis,structure and in vitro biological activity[J]. RSC Advances,2014,4(4):2004-2022.
[12]Tan C,Lai S,Wu S,et al. Nuclear permeable ruthenium(II)β-carboline complexes induce autophagy to antagonize mitochondrial-mediated apoptosis[J].J Med Chem,2010,53(21):7613-7624.
[13]Cardoso CR,Lima MVS,Cheleski J,et al. Luminescent ruthenium complexes for theranostic applications[J]. J Med Chem,2014,57(11):4906-4915.
[14]赵彦超,顾耘.细胞凋亡通路研究进展[J].现代医学,2013,5(23):490-492.
[15]谭生权,王璋.白藜芦醇调节VEGF/Wnt/β-catenin通路对食管癌细胞的抑制作用[J].解剖学研究,2018,40(2):108-110.
[16]华夏,李广兴,刘仲涛,等.人参皂苷Rh2下调Wnt/β-catenin信号通路调控SHG44细胞的增殖和凋亡[J].解剖学研究,2018,40(3):189-192.